Project 4 - Abstract The principle that adoptively transferred T lymphocytes have therapeutic promise for HIV infection is well established. Our long range goals are to engineer T cells so that they can control HIV-1 replication in the absence of HAART. Our companies? scientific founders have pioneered the use of cell and gene therapy to treat HIV-1, and we will use this expertise to better study the relationship between T cells that have been rendered resistant to HIV-1 infection and specific for HIV to control of HIV-1 replication in the absence of ART. Our project serves as a main integration site for this U19 consortium as the most promising approaches generated by Projects 1-3 will be incorporated into Phase I clinical trial that we design, execute and analysis. Additionally, we will develop a commercial T cell manufacturing platform that will give HIV infected individuals access to Chimeric Antigen Receptor (CAR) therapy. The specific aims of this Project are: 1) Define the optimal method to expand T cells for HIV cure studies: Using Tmunity?s T cell expansion expertise and proprietary technology, we will systematically address the best media, artificial APC, and manufacturing platform to develop an optimized protocol to expand highly functional T cells with superior engraftment potential to be used as part of HIV cure regimens. 2) Design and implement a Phase I clinical trial to test the ability of engineered T cells to prevent viral rebound during an analytical treatment interruption: With input from the members of this U19, scientific advisory board (SAB) and NIH program officers, we will plan and execute a Phase I clinical trial that uses this new manufacturing protocol established in Aim 1. 3) Perform correlative studies on samples collected in Aim 2 to develop testable hypotheses that could improve adoptive T cell therapy for HIV infection: Using banked cells collected from informative time points, we will perform validated assays that examine T cell function and persistence and the ability of the chosen intervention to control HIV replication in the absence of ART.